Osteoinductive and Osteogenic Capacity of Freeze-Dried Bovine Bone Compared to Deproteinized Bovine Bone Mineral Scaffold in Human Umbilical Cord Mesenchymal Stem Cell Culture: An In Vitro Study

Abstract

Objective Freeze-dried bovine bone scaffold (FDBB) or decellularized FDBB (dc-FDBB) was developed as an ideal scaffold with osteoinductive properties. This research aims to compare the osteoinductive properties marked by the expression of runt-related transcription factor-2 (RUNX2) and Osterix (OSX) and the osteogenic capacity of these scaffolds imbued with human umbilical cord mesenchymal stem cells (hUCMSCs). Materials and Methods This study was performed in five experimental groups: a negative control group (C-) of hUCMSCs with a normal growth medium, a positive control group (C + ) of hUCMSCs with an osteogenic medium, experimental group 1 (E1) with an FDBB conditioned medium (CM), and experimental group 2 (E2) with a dc-FDBB-CM, and a third experimental group (E3) consisting of a DBBM-CM. Alizarin red staining was performed to qualitatively assess osteoinductive capacity. RUNX2 and OSX expression was quantified using real-time quantification polymerase chain reaction with two replications on day six (D6) and day 12 (D12) as fold changes. Results This experiment revealed that hUCMSCs were positively expressed by CD73, CD90, and CD105 but were not expressed by CD34. Alizarin red staining showed that E1 had the most calcium deposition on D6 and D12, followed by E3 and then E2 The RUNX2 and OSX expression was higher in E1 but this difference was not significant. The OSX expression in E1,E2,E3 was lower on D12 and C+ of OSX had the highest expression. There was a significant difference of fold change measured between all groups (p < 0.05), and there was no significant difference between any of the groups treated with OSX and RUNX2 on D6 and D12. Conclusion FDBB osteoinduction and osteogenic capacity were higher when compared with DBBM and dc-FDBB.

Preosteoblast Adhesion and Viability Study of Freeze-Dried Bovine Bone Block Scaffold Coated with Human Umbilical Cord Mesenchymal Stem Cell Secretome

“…Therefore, the osteoinduction properties of FDBB scaffolds are superior to those of dcFDBB scaffolds because they are supported by suitable organic components. 8 However, the observations at 24 hours differed. The DBBM-CM scaffold showed the highest average number of adherent cells, but the differences were not statistically significant.…”

“…The low cell viability on dcFDBB-CM scaffolds was also caused by the low cell adhesion owing to destruction of the organic materials. 8 The high cell viability on the DBBM-CM scaffold was supported by direct cell adhesion to the scaffold. The direct adhesion process is supported by the surface topography characteristics, such as pore size and degree of roughness.…”

“…3,20 FDBB has osteoinductive and osteodifferentiation potential because it retains the organic components, growth factors, glycosaminoglycans, noncollagenous ECM proteins, and scaffold morphology. 8,21 Cell adhesion is a complex dynamic process involving the adsorption of proteins onto surfaces and expression of specific peptide sequences. Cells attach to certain surfaces via integrins and usually die if they fail to attach.…”

“…The heating and chemical extraction processes used to remove all organic components result in deproteinized bovine bone mineral (DBBM), the freeze-drying process results in freeze-dried bovine bone (FDBB), and the addition of the decellularization process results in decellularized FDBB (dcFDBB). [6][7][8] The scaffolds in BTE must be three-dimensional (3D) to support cell growth. These 3D scaffolds can provide a specific architecture, namely, a matrix that provides temporary mechanical support for cell migration, proliferation, and differentiation.…”

The diagnostic challenges and two-step surgical approach to an infected dentigerous cyst resembling a unicystic ameloblastoma: A case report

“…Radiographic images such as panoramic radiographs could highlight the anatomical position of the pathologic lesion, but they also present disadvantages because many lesions have a similar radiographic appearance. 20 A biopsy is the gold standard for obtaining a microscopic image through a histopathologic examination, as conducted in this case. To determine the diagnosis, a triple analysis should be performed based on clinical signs, radiographic images, and histopathology.…”